Study On The Association Of Environmental Factors,Antioxidant Nutrients And Inflammatory Gene Polymorphisms With Oral Cancer Risk And Its Prognosis

ObjectiveTo develop environmental exposure indexes and evaluate the effects of environmental factors on oral cancer in men and women;to assess the association between antioxidant nutrients,inflammatory gene polymorphisms and oral cancer risk;to explore the potential biological function of SNPs in NFKB1 gene promoter on oral cancer;to develop and validate a prognostic nomogram for patients with oral cancer.Methods1.A case-control study was performed including 908 oral cancer patients and 2542 controls.Epidemiologic data were collected through face-to-face interviews using a structure questionnaire.Unconditional logistic regression model was used to assess the effects of environmental factors on oral cancer in men and women.Environmental exposure indexes were developed and then evaluated by receiver operating characteristic curve.2.Dietary antioxidant nutrients intakes were estimated using food frequency questionnaires(FFQ)and were adjusted for total energy by residual method.The effects of antioxidant nutrients were analyzed using unconditional logistic regression models.The association between serum folic acid and oral cancer risk was further validated in the validation group.3.3ml of venous blood was collected for DNA extraction,and 26 SNPs on inflammatory signaling pathways were genotyped by Mass ARRAY platform.The association of SNPs with oral cancer and gene-environment interactions were analyzed by unconditional logistic regression and crossover analysis.4.NFKB1 promoter luciferase reporter plasmids containing rs28362491 Ins/Del and rs72696119 C/G were constructed.The effect of different haplotypes on the transcriptional activity was estimated by dual-luciferase reporter assay,and the binding of transcription factor was detected by electrophoretic mobility shift assay.5.A prospective study involving 908 oral cancer patients was conducted to collect clinical data and follow-up data.Multivariate Cox regression model was used to assess independent predictive factors in oral cancer prognosis.Nomogram was established by R software.The discrimination ability of nomogram was assessed by Harrell's C-index.Results1.Specific risk factors for male oral cancer included smoking pack-year?20,average daily alcohol consumption ?20g,duration of denture ?10 years;while particular protective factors included intake of fish ?3 times/week,seafood?1 times/week,milk and dairy products ?1 times/week.Specific risk factors for female oral cancer are passive smoking,cooking oil fume exposure and age of first intercourse <20;while particular protective factor was intake of beans and soy products? 1 times/week.The higher the environmental exposure index,the higher the risk of oral cancer was observed.The index also has a high predictive ability for the risk of oral cancer.2.High dietary intake of vitamin C,vitamin E,folic acid and ?-carotene were associated with decreased risk of oral cancer.Serum folic acid was inversely associated with oral cancer risk.There was negative multiplicative and additive interaction between serum folic acid and smoking or alcohol consumption in the validation group.3.NFKB1 rs28362491 Del/Del and rs72696119 GG genotypes were associated with increased risk of oral cancer.There was a strong linkage disequilibrium between rs28362491 and rs72696119(r2=0.971).Compared with the IC haplotype,individuals with DG haplotype had a 1.25-fold increased risk of oral cancer(OR=1.25,95% CI: 1.02-1.53).Additionally,IL1 B rs1143627 GG genotype were significantly associated with a decreased risk of oral cancer.4.Tca-8113 or CAL-27 cells transfected with IG or DC or DG plasmids showed much lower luciferase activity than that of cells transfected with IC plasmid.The results of EMSA showed that Ins probe had strong binding activity to nuclear protein,and the mutant Del probe showed no binding.Bioinformatics software predicted that there was an alpha-CP1 binding sites only for rs28362491 Ins/Ins,but not for rs28362491 Del/Del.5.TNM stage,tumor differentiation,surgical treatment,radiotherapy and NLR before treatment were the independent prognostic factors for patients with oral cancer.The results of the training and validation sets all demonstrated that the discrimination of the nomogram was superior to AJCC staging system(the 7th edition).Conclusions1.There are gender differences in the environmental factors of oral cancer.The environmental exposure indexes of male and female have high predictive ability and application value.2.High dietary intake of vitamin C,vitamin E,folic acid and ?-carotene are the protective factors of oral cancer.3.NFKB1 rs28362491,rs72696119 and IL1 B rs1143627 were associated with the susceptibility of oral cancer.4.NFKB1 rs28362491 and rs72696119 polymorphisms can affect the binding ability of transcription factor and reduce the expression of NF-?B1,thereby affecting the occurrence and development of oral cancer.5.Nomogram prediction model was successfully established and validated to predict the overall survival of oral cancer patients,with high prediction accuracy and clinical application value.